Detection of apoptosis
The TUNEL (terminal deoxynucleotidyl transferase-mediated
dUTP nick end labeling) assay was used to assess cardiomyocyte
apoptosis, according to the instructions in the reagent kits
(Roche Molecular Biochemicals). These sections were visualized
by an Olympus visible light microscope at 400×
magnification.
TUNEL-positive cardiomyocytes were carefully evaluated under
double-blind conditions. In each visual field, the number of positive
cell nuclei was calculated among 200 myocardial cell nuclei.
The rate of apoptotic cell nuclei is defined as apoptotic positive cell
nuclei/total cell nuclei in the field
×
100.